Acute oral toxicity and anti-inflammatory evaluation of methanolic extract of Rotula aquatica roots in Wistar rats

Background: The plant Rotula aquatica Lour. was traditionally well known due to its large number ofpharmacological action and medicinal uses. The plant is a necessary component of many Ayurvedic drugpreparations since historical times. It is widely used as a crucial ancient drug for kidney and bladderstones.Objectives: The main objective of the study was to evaluate the acute toxicity and anti inflammatoryefficacy of methanolic extract of R. aquatica Lour. in in vivo models.Materials and methods: The qualitative phytochemical analysis and invitro antioxidant activity of theroots of methanolic extract of R. aquatica Lour. (MERA) was evaluated. The acute toxicity effect of MERAwas evaluated with two different doses (550, 2000 mg/kg body weight), were administrated orally toWistar rats. The rats were observed for sign and symptoms of toxicity and mortality for 14 days. Theparameters measured including relative organ weight, blood, biochemical and histopathological parameters of hepatic and renal toxicity. The anti-inflammatory effect of MERA was also evaluated incarrageenan and dextran-induced paw edema models.Results: The phytochemical evaluation of MERA shows the presence of secondary metabolites like alkaloids, flavonoids, phenolics and tannins, phytosterols, reducing sugars, proteins and terpenoids. Theresults of in-vitro antioxidant evaluation of MERA reveal its capability to scavenging free radical at alower concentration. The MERA did not show any visible signs of toxicity up to the dose of 2000 mg/kgbody weight. The results obtained from our carrageenan and dextran-induced paw edema model studyalso proved the anti-inflammatory effect of MERA in rat model.Conclusion: The result shows the potential of MERA as an anti-inflammatory drug to reduce the signs ofinflammation devoid of any toxic effect.© 2018 The Authors. Published by Elsevier B.V. on behalf of Institute of Transdisciplinary Health Sciencesand Technology and World Ayurveda Foundation. This is an open access article under the CC BY-NC-NDlicense (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Rotula aquatica Lour. inhibits growth and biofilm formation of clinically isolated uropathogenic Escherichia coli

Objective: To evaluate the anti-bacteria! and anti-biofilm activity of ethyl acetate fraction of Rotula aquatica Lour. (EFRA) against clinically isolated uropathogenic Escherichia coli. Methods: In vitro antibacterial and anti-biofilm studies were employed. The antimicrobial activity of EFRA was assayed by the well diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the active fraction were determined by Resazurin method. The time-kill kinetic assay, acridine orange-ethidium bromide staining, propidium iodide uptake assay, and scanning electron microscopic (SEM) analysis were done to evaluate the efficacy of EFRA in killing uropathogenic Escherichia coli. The anti-biofilm activity was determined by 3-[4,5- dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium-bromide (MTT) assay and specific biofilm formation assay. Results: The well diffusion assay of EFRA showed a very clear zone of inhibition against Escherichia coli BRL-17. The MIC and MBC of EFRA were 2.5 mg/mL and 5 mg/mL, respectively. The time-kill kinetic assay, fluorescence microscopic analysis, propidium iodide uptake assay, and SEM analysis displayed the effect of EFRA in killing the bacteria. The MTT assay and specific biofilm formation assay showed that EFRA prevented the formation of biofilms. Conclusions: The results of the present study confirm that EFRA could prevent bacterial growth and inhibit its biofilm formation.

Rotula aquatica Lour. inhibits growth and biofilm formation of clinically isolated uropathogenic Escherichia coli

Objective: To evaluate the anti-bacterial and anti-biofilm activity of ethyl acetate fraction of Rotula aquatica Lour. (EFRA) against clinically isolated uropathogenic Escherichia coli. Methods: In vitro antibacterial and anti-biofilm studies were employed. The antimicrobial activity of EFRA was assayed by the well diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the active fraction were determined by Resazurin method. The time-kill kinetic assay, acridine orange-ethidium bromide staining, propidium iodide uptake assay, and scanning electron microscopic (SEM) analysis were done to evaluate the efficacy of EFRA in killing uropathogenic Escherichia coli. The anti-biofilm activity was determined by 3-[4,5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium-bromide (MTT) assay and specific biofilm formation assay. Results: The well diffusion assay of EFRA showed a very clear zone of inhibition against Escherichia coli BRL-17. The MIC and MBC of EFRA were 2.5 mg/mL and 5 mg/mL, respectively. The time-kill kinetic assay, fluorescence microscopic analysis, propidium iodide uptake assay, and SEM analysis displayed the effect of EFRA in killing the bacteria. The MTT assay and specific biofilm formation assay showed that EFRA prevented the formation of biofilms. Conclusions: The results of the present study confirm that EFRA could prevent bacterial growth and inhibit its biofilm formation.

Total Phenolics and Antioxidant Potentials of Rotula aquatica Lour.

Rotula aquatica Lour is a small branched shrub that grows characteristically among rocks and trailing over gravel in stream beds. It belongs to the family Boraginaceae and widely distributed throughout India. It is commonly called as Pasanabheda. The present study was aimed to evaluate the total phenolics and in vitro antioxidant activity in root and leaf extracts of Rotula aquatica Lour. The dried roots and leaves were subjected to soxhlet extraction utilizing chloroform and methanol followed by preliminary qualitative phytochemical analysis. Total phenolic content was estimated by Folin-Ciocalteu method and in vitro antioxidant assays were performed using various radical scavenging activities viz. DPPH, superoxide anion, nitric oxide, hydroxyl and iron chelating activities at different concentrations. The results of the total phenolic content estimation was expressed in terms of equivalence with standard and the antioxidant potentiality of various scavenging and chelating activity were expressed in terms of IC50 values. The results showed that both root and leaves possess significant antioxidant activity even though the former was found to be superior than the latter in terms of parameters assessed. Further the methanol extracts were proved to be an ideal for the materials subjected to investigation.

Antilithiatic activity of poly-herbal formulation tablets by in-vitro method.

Hydroalcoholic extracts of Kalanchoe pinnata and Rotula aquatica and its combination were formulated herbal tablets, evaluated for antilithiatic in vitro method. The homogenous precipitation method was used. The study was carried out in glass tubes. The buffer system used was TRIS buffer pH 7.4. The experiment consists of the following tubes for control and test, 25 ml each of 25 mM CaCl2. 2H2O, 25 mM Na2HPO4. 2H2O or 25mM Na2C2O4. To the tubes of each set, tablet formulation or an equal amount of vehicle was added. The tubes were incubated at 37°C for 4 h. The precipitate of calcium phosphate was generated by mixing 1 ml of solution from the tubes having calcium chloride dihydrate and disodium hydrogen phosphate monohydrate and Calcium oxalate precipitate was generated by mixing 1 ml of solutions from the tubes having calcium chloride dihydrate and sodium oxalate solutions. Calcium was estimated using titrimetry and phosphorus was estimated using colorimetric analysis. Appropriate standard curves were done with each set of experiments. The amounts of precipitate of calcium and phosphate were determined in each of the respectively. The percent inhibition of the test was calculated in comparison with the control samples. Herbal tablet formulation showed antilithiatic activity to the marketed formulation in terms of inhibiting the formation of phosphate precipitate but showed a significantly better potential in preventing the formation of the calcium precipitate. The herbal tablet formulation of Kalanchoe pinnata and Rotula aquatica have inhibitory effect on calcium oxalate crystallization thus may be beneficial in the treatment of renal lithiasis.

Evaluation of Physiochemical Screening and Standardization on the Root of Rotula aquatica Lour.

The root of Rotula aquatica is also called as pashanbed, belonging to the family Borogenaceae. It is widely distributed in India from kumaun to Assam and western to southern India.The medicinal values of plant lie in their component phytochemicals such as alkaloids, flavonoids, phenolic compounds and other nutrients like as amino acid, proteins, which produce a definite physiological action on the human body. The proximate values such as moisture content, ash value, extractive value, were carried out. The dried roots powder was subjected to successive extraction with different solvents like petroleum ether, benzene, chloroform, methanol, ethanol and water. Macroscopic as well as microscopic studies of any crude drug are the primary steps to establish its botanical quality control before going to other studies. Hence Standardization of crude drug plays a very important role in identifying the purity and quality of crude drugs. The present investigation reveals Standardization which include Moisture Content, Total ash, Acid insoluble ash, Water soluble ash, Water soluble Extractive value, Alcohol soluble Extractive value, Phytochemical screening, and further isolation and identification of Phytoconstituents.

Growth inhibition of struvite crystals by the aqueous root extract of Rotula aquatica.

Formation of urinary stone is a serious and debilitating problem throughout the world. In the present study, the inhibitory effect of aqueous extract of root of Rotula aquatica was investigated against struvite crystals (one of the components of urinary stone) grown in vitro using single diffusion gel growth technique. For setting the gel, sodium metasilicate solution (specific gravity 1.05) and 0.5 M aqueous solution of ammonium dihydrogen phosphate were mixed, so that the pH of the mixture could be set at 7.0. Equal amounts of supernatant solution of magnesium acetate (1.0 M) prepared with 0.0%, 0.5% and 1% concentrations of the extract were gently poured on the set gels. It was observed that the number, dimension, total mass, total volume, growth rate and depth of growth of struvite crystals decreased with the increasing extract concentrations in the supernatant solutions. The enhancement of dissolution rate and fragmentation of struvite crystals suggested potential application of the extract for inhibition of struvite type urinary stone.